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Droegemueller, C.

Publications and source records attributed to Droegemueller, C..

3 recordsLinked to original sources

An endogenous retrovirus insertion disrupting bovine ALKBH8 causes a failure-to-thrive syndrome with immunodeficiency associated with juvenile mortality in Brown Swiss cattle

The Brown Swiss (BS) cattle breed is one of the major Swiss dairy breeds. Intensive selection and the widespread use of few elite sires in artificial insemination have increased inbreeding and the occurrence of deleterious recessive alleles in the homozygous state. Analyzing life trajectories in large, genotyped cohorts can identify hidden recessive disorders that are difficult to detect using traditional case-control association testing. Long-read DNA sequencing enables precise detection of causal alleles, including structural variants. This study aimed to (1) identify cryptic recessive loci affecting rearing performance in Swiss BS cattle, (2) evaluate their impact on survival, (3) characterize the associated phenotype, (4) identify the causal variant using long-read whole-genome sequencing, and (5) assess its functional impact. Using Homozygous Haplotype Enrichment/Depletion (HHED) mapping, we identified a risk haplotype (BH39) on chromosome 15 spanning from 16,276,819 bp to 16,446,984 bp that was associated with increased juvenile mortality within the first 180 days of life when present in the homozygous state. The BH39 occurred at a frequency of approximately 4.5% in Swiss BS cattle and 5.3% in German and Austrian BS cattle, and homozygous carriers exhibited a significantly reduced first-year survival rate. Five females homozygous for BH39 underwent clinical examination. They all showed recurrent respiratory disease, impaired growth, poor body condition, rough hair coat, and brown-discolored teeth. Pathological examination revealed bronchopneumonia and eosinophilic enteritis. Clinicopathological findings indicated failure to thrive and immunodeficiency. Long-read WGS of two BH39 homozygous calves revealed a private homozygous coding variant that was in high linkage disequilibrium with BH39. The identified structural variant was an insertion of a large transposable element (10.4 kb ERVK[2-1-LTR]) into the third exon of ALKBH8 (NM_001080341.2 c.267_268indel). Full-length RNA sequencing of cerebellum and liver from a homozygous calf revealed that the endogenous retrovirus (ERV) insertion introduces a cryptic transcription termination signal, truncating ALKBH8 mRNA. This study demonstrates that exploring population-scale genomic data and mining thousands of life-history records, followed by veterinary follow-up evaluations and molecular genetic analyses, provides an effective strategy for identifying cryptic recessive disorders that shorten the lifespan of cattle. The findings provide strong evidence that the ERV insertion into the coding sequence of ALKBH8 represents a loss-of-function variant that causes a previously undescribed recessive disorder that results in increased rearing loss. Interpretive summaryWe identified a recessive disorder in Brown Swiss cattle that causes retarded growth, recurrent infections, immunodeficiency, and increased mortality during the first year of life. Using population-scale genomic data, clinical investigations, and long-read sequencing, we linked the disorder to an exonic transposable element insertion disrupting ALKBH8. The identification of the causal variant now enables direct genetic testing and the implementation of genome-based mating strategies to avoid carrier-by-carrier matings and, consequently, prevent the birth of affected homozygous offspring. We demonstrate the utility of integrating large-scale breeding records, veterinary phenotyping, and advanced genomics to identify hidden defects affecting livestock health and productivity.

genomics↗

Taurine pangenome uncovers a segmental duplication upstream of KIT associated with depigmentation in white-headed cattle

Cattle have been selectively bred for coat color, spotting, and depigmentation patterns. The assumed autosomal dominant inherited genetic variants underlying the characteristic white head of Fleckvieh, Simmental, and Hereford cattle have not been identified yet, although the contribution of structural variation upstream the KIT gene has been proposed. Here, we construct a graph pangenome from 24 haplotype assemblies representing seven taurine cattle breeds to identify and characterize the white head-associated locus for the first time based on long-read sequencing data and pangenome analyses. We introduce a pangenome-wide association mapping approach which examines assembly path similarities within the graph to reveal an association between two most likely serial alleles of a complex structural variant 66 kb upstream KIT and facial depigmentation. The complex structural variant contains a variable number of tandemly duplicated 14.3 kb repeats, consisting of LTRs, LINEs, and other repetitive elements, leading to misleading alignments of short and long reads when using a linear reference. We align 250 short-read sequencing samples spanning 15 cattle breeds to the pangenome graph, further validating that the alleles of the structural variant segregate with head depigmentation. We estimate an increased count of repeats in Hereford relative to Simmental and other white-headed cattle breeds from the graph alignment coverage, suggesting a large under-assembly in the current Hereford-based cattle reference genome which had fewer copies. We show that exploiting assembly path similarities within graph pangenomes can reveal trait-associated complex structural variants.

genomics↗

A living biobank of canine mammary tumor organoids as a comparative model for human breast cancer

Mammary tumors in dogs hold great potential as naturally occurring breast cancer models in translational oncology, as they share the same environmental risk factors, key histological features, hormone receptor expression patterns, prognostic factors, and genetic characteristics as their human counterparts. We aimed to develop in vitro tools that allow functional analysis of canine mammary tumors (CMT), as we have a poor understanding of the underlying biology that drives the growth of these heterogeneous tumors. We established the long-term culture of 24 organoid lines from 16 patients, including organoids derived from normal mammary epithelium or benign lesions. CMT organoids recapitulated key morphological and immunohistological features of the primary tissue from which they were derived, including hormone receptor status. Furthermore, genetic characteristics (driver gene mutations, DNA copy number variations, and single-nucleotide variants) were conserved within tumororganoid pairs. We show how CMT organoids are a suitable model for in vitro drug assays and can be used to investigate whether specific mutations predict therapy outcomes. In addition, we could genetically modify the CMT organoids and use them to perform pooled CRISPR/Cas9 screening, where library representation was accurately maintained. In summary, we present a robust 3D in vitro preclinical model that can be used in translational research, where organoids from normal, benign as well as malignant mammary tissues can be propagated from the same patient to study tumorigenesis.

cancer biology↗